本文選題：膠質(zhì)瘤 + 能量代謝　； 參考：《浙江大學(xué)》2014年博士論文

【摘要】：一基于膠質(zhì)瘤代謝網(wǎng)絡(luò)的小分子化合物抗腫瘤作用及機(jī)制研究目的：膠質(zhì)瘤是一類常見(jiàn)的原發(fā)性顱內(nèi)腫瘤,發(fā)病率約占原發(fā)性惡性腦腫瘤的50%以上。盡管現(xiàn)代醫(yī)學(xué)技術(shù)有了極大發(fā)展,但是膠質(zhì)瘤患者平均生存時(shí)間不超過(guò)15個(gè)月,而且手術(shù)過(guò)后易復(fù)發(fā),現(xiàn)在仍然沒(méi)有理想的藥物及治療方法。有氧糖酵解是腫瘤細(xì)胞主要的能量代謝方式,本實(shí)驗(yàn)主要研究受試藥物ZZZ-1針對(duì)膠質(zhì)瘤細(xì)胞代謝網(wǎng)絡(luò)的抗腫瘤作用及其機(jī)制。實(shí)驗(yàn)設(shè)計(jì)：本研究用SRB法及集落形成實(shí)驗(yàn)檢測(cè)受試藥物ZZZ-1對(duì)一系列膠質(zhì)瘤細(xì)胞株的抗腫瘤藥效,研究其對(duì)膠質(zhì)瘤細(xì)胞能量代謝過(guò)程中葡萄糖消耗、丙酮酸消耗、乳酸產(chǎn)生、細(xì)胞內(nèi)ATP的產(chǎn)生及氧氣消耗比例(OCR)的影響,進(jìn)一步研究藥物對(duì)膠質(zhì)瘤細(xì)胞線粒體膜電位ΔΨm、ROS產(chǎn)生、細(xì)胞衰老、細(xì)胞凋亡及其相關(guān)蛋白、干細(xì)胞標(biāo)志Bmi-1、正常星形膠質(zhì)細(xì)胞標(biāo)志GFAP及糖酵解過(guò)程中關(guān)鍵蛋白PFKFB3、LDH5表達(dá)的影響。用氧化磷酸化過(guò)程中ATP合成酶抑制劑寡霉素研究ZZZ-1的潛在作用機(jī)制。本實(shí)驗(yàn)進(jìn)一步研究ZZZ-1在動(dòng)物體內(nèi)實(shí)驗(yàn)中對(duì)膠質(zhì)瘤增殖抑制、血管內(nèi)皮生長(zhǎng)因子VEGF表達(dá)、糖酵解過(guò)程中關(guān)鍵酶、干細(xì)胞指標(biāo)和分化指標(biāo)的影響。實(shí)驗(yàn)結(jié)果：1)ZZZ-1具有強(qiáng)效的抗膠質(zhì)瘤藥效學(xué)作用：可顯著抑制人及大鼠膠質(zhì)瘤細(xì)胞增殖,抑制膠質(zhì)瘤細(xì)胞集落形成,并良好的呈劑量依賴性；2)誘導(dǎo)膠質(zhì)瘤細(xì)胞死亡及分化：誘導(dǎo)膠質(zhì)瘤細(xì)胞AΨm去極化及ROS過(guò)度產(chǎn)生,抑制PCNA、 survivin的表達(dá),誘導(dǎo)大鼠膠質(zhì)瘤C6細(xì)胞凋亡及人膠質(zhì)瘤細(xì)胞U-87MG細(xì)胞衰老,抑制干細(xì)胞指標(biāo)Bmi-1的表達(dá),促進(jìn)膠紙瘤細(xì)胞向正常星形膠質(zhì)細(xì)胞分化；3)誘導(dǎo)膠質(zhì)瘤細(xì)胞能量代謝方式發(fā)生改變：ZZZ-1處理膠質(zhì)瘤細(xì)胞24 h可誘導(dǎo)細(xì)胞葡萄糖及丙酮酸消耗及乳酸產(chǎn)生增多,可顯著提高細(xì)胞內(nèi)ATP產(chǎn)生量和氧氣消耗量,藥物處理細(xì)胞72 h可降低糖酵解過(guò)程中關(guān)鍵蛋白PFKFB3、LDH5的表達(dá),寡霉素可降低ZZZ-1引起的膠質(zhì)瘤細(xì)胞增殖抑制,抑制△Ψm的去極化及ROS的產(chǎn)生,減少葡萄糖的消耗及ATP的產(chǎn)生；ZZZ-1可減少寡霉素引起的代償性乳酸的產(chǎn)生；4)抑制裸小鼠移植瘤生長(zhǎng)：在U-87MG裸小鼠移植瘤模型中,ZZZ-1可選擇性的抑制腫瘤細(xì)胞的生長(zhǎng),并且動(dòng)物體重?zé)o降低,具有很高的安全性；可降低PCNA. survivin及糖酵解過(guò)程中關(guān)鍵蛋白PFKFB3、LDH5的表達(dá)；促進(jìn)膠質(zhì)瘤細(xì)胞向正常星形膠質(zhì)細(xì)胞、神經(jīng)細(xì)胞及小膠質(zhì)細(xì)胞分化。結(jié)論：結(jié)果提示,ZZZ-1對(duì)膠質(zhì)瘤細(xì)胞具有顯著而穩(wěn)定的抗腫瘤作用,可通過(guò)多層次、多途徑調(diào)控膠質(zhì)瘤細(xì)胞能量代謝、誘導(dǎo)細(xì)胞凋亡及衰老、促進(jìn)膠質(zhì)瘤細(xì)胞向正常神經(jīng)細(xì)胞、星形膠質(zhì)細(xì)胞及小膠質(zhì)細(xì)胞分化等方面起到抑制膠質(zhì)瘤細(xì)胞增殖的作用。二小分子化合物CADPE對(duì)腸癌多靶點(diǎn)調(diào)控作用及機(jī)制研究目的：結(jié)直腸癌是一類嚴(yán)重影響全球人類健康的疾病,每年世界范圍內(nèi)大約有一百萬(wàn)人被診斷出患有腸癌,同年有超過(guò)五十萬(wàn)人死于腸癌。本實(shí)驗(yàn)主要研究咖啡酸3,4-二羥基苯乙酯(CADPE)對(duì)結(jié)直腸癌的抗腫瘤藥效、作用機(jī)制及可能的作用靶點(diǎn)。實(shí)驗(yàn)設(shè)計(jì)：本研究用SRB法及集落形成實(shí)驗(yàn)檢測(cè)CADPE對(duì)四株人結(jié)直腸癌細(xì)胞的抗腫瘤藥效,并研究CADPE對(duì)腸癌細(xì)胞的細(xì)胞周期、凋亡及周期和凋亡相關(guān)蛋白的影響。用MAPK信號(hào)通路的四種抑制劑研究CADPE對(duì)腸癌細(xì)胞的潛在作用機(jī)制。本實(shí)驗(yàn)進(jìn)一步在體內(nèi)動(dòng)物實(shí)驗(yàn)中研究CADPE對(duì)腸癌的增殖抑制、細(xì)胞凋亡及細(xì)胞周期相關(guān)蛋白的影響,并評(píng)估CADPE對(duì)人正常腸癌組織細(xì)胞的影響。實(shí)驗(yàn)結(jié)果：本研究發(fā)現(xiàn)CADPE可顯著抑制人結(jié)直腸癌細(xì)胞增殖和集落形成能力,并誘導(dǎo)腸癌細(xì)胞G0/G1期細(xì)胞周期阻滯和細(xì)胞凋亡；CADPE對(duì)腸癌的抑制作用與p38 MAPK、p53、p21及p16等信號(hào)通路的激活、核轉(zhuǎn)錄因子κB及其下游靶基因信號(hào)轉(zhuǎn)導(dǎo)子和轉(zhuǎn)錄激活因子3 (STAT3)和c-Myc的抑制有關(guān)；CADPE還可以下調(diào)生存素survivin及細(xì)胞周期相關(guān)蛋白(如cyclin D1、CDK4、CDK6、p-Rb和E2F-1)的表達(dá)；P38 MAPK的抑制劑可減少CADPE引起的細(xì)胞增殖抑制、細(xì)胞周期阻滯及細(xì)胞凋亡,但是ERK和JNK抑制劑沒(méi)有此類作用。在SW620裸小鼠移植瘤模型中,CADPE同樣可抑制腫瘤生長(zhǎng)及誘導(dǎo)細(xì)胞凋亡,并降低PCNA、survivin及細(xì)胞周期相關(guān)蛋白(cyclin D1、CDK4、CDK6、p-Rb)的表達(dá),并且抗腫瘤作用濃度范圍內(nèi)的CADPE對(duì)正常細(xì)胞無(wú)細(xì)胞毒作用,在體內(nèi)及體外實(shí)驗(yàn)中均具有很高的安全性。結(jié)論：實(shí)驗(yàn)結(jié)果提示CADPE在調(diào)控結(jié)直腸癌細(xì)胞增殖方面起著重要的作用,并可通過(guò)多靶點(diǎn)進(jìn)行調(diào)控,從而抑制腸癌細(xì)胞增殖、引起腸癌細(xì)胞周期阻滯、誘導(dǎo)腸癌細(xì)胞凋亡。
[Abstract]:A study on the anti-tumor effect and mechanism of small molecular compounds based on the metabolic network of glioma: glioma is a common type of primary intracranial tumor, which accounts for more than 50% of the primary malignant brain tumors. Although modern medical technology has developed greatly, the average survival time of the patients with glioma is not more than 15 months. It is easy to relapse after the operation, and there is still no ideal medicine and treatment. Aerobic glycolysis is the main energy metabolism mode of tumor cells. This experiment mainly studies the anti-tumor effect and mechanism of drug ZZZ-1 on the metabolic network of glioma cells. Experimental design: This study was tested by SRB method and colony formation test. The antitumor effect of drug ZZZ-1 on a series of glioma cell lines, and the effects of glucose consumption, pyruvate consumption, lactic acid production, production of intracellular ATP and oxygen consumption ratio (OCR) on the energy metabolism of glioma cells, and further research on the mitochondrial membrane potential of glioma cells, delta m, ROS production, cell senescence, and cell aging Apoptosis and its related proteins, stem cell marker Bmi-1, normal astrocyte marker GFAP and the influence of the key protein PFKFB3, LDH5 expression in glycolysis process. The potential mechanism of ZZZ-1 was studied by oligomycin, a ATP synthetase inhibitor in the process of oxidative phosphorylation. This experiment studied the glioma of ZZZ-1 in animal experiments. Proliferation inhibition, expression of vascular endothelial growth factor VEGF, the influence of key enzyme, stem cell index and differentiation index in glycolysis. Experimental results: 1) ZZZ-1 has a potent anti glioma pharmacodynamic effect: it can significantly inhibit the proliferation of glioma cells in human and rat, inhibit glioma cell colony formation, and have a good dose dependence; 2) induced death and differentiation of glioma cells: inducing glioma cell A m depolarization and ROS overproduction, inhibiting the expression of PCNA, survivin, inducing apoptosis of C6 cells in glioma and senescence of human glioma cell U-87MG cells, inhibiting the expression of stem cell index Bmi-1, promoting the differentiation of gelatinous tumor cells to normal astrocytes; 3) lure. Changes in the energy metabolism of glioma cells: ZZZ-1 treatment of glioma cells 24 h can induce the consumption of glucose and pyruvic acid and the increase of lactic acid, which can significantly increase the amount of intracellular ATP production and oxygen consumption. Drug treatment cells 72 h can reduce the expression of PFKFB3, LDH5, the key protein in glycolysis. Oligomycin can be reduced. Low ZZZ-1 induced proliferation inhibition of glioma cells, depolarization of delta m depolarization and ROS production, reducing glucose consumption and ATP production; ZZZ-1 can reduce the production of compensatory lactic acid induced by oligomycin; 4) inhibit the growth of xenografts in nude mice: ZZZ-1 can selectively inhibit tumor cells in the nude mice model of U-87MG Growth, and the animal weight has no decrease, and has high safety; it can reduce the expression of PFKFB3, LDH5, the key protein in PCNA. survivin and glycolysis; promote the differentiation of glioma cells to normal astrocytes, nerve cells and microglia. Conclusion: the results suggest that ZZZ-1 has a significant and stable resistance to glioma cells. Tumor action can regulate the energy metabolism of glioma cells by multilevel and multiple pathways, induce apoptosis and senescence, and promote the inhibition of glioma cells to normal nerve cells, astrocytes and microglia differentiation. Two small molecule compound CADPE regulates the multiple target of colorectal cancer. Objective: colorectal cancer is a kind of disease that seriously affects human health worldwide. About one million people are diagnosed with colorectal cancer worldwide every year. More than five hundred thousand people died of cancer in the same year. This experiment mainly studied the antitumor effect of caffeic acid 3,4- two hydroxybenzene ethyl ester (CADPE) on colorectal cancer. Possible target targets. Experimental design: This study detected the antitumor efficacy of CADPE on four human colorectal cancer cells by SRB method and colony formation test, and studied the effect of CADPE on the cell cycle, apoptosis, cycle and apoptosis related proteins of colorectal cancer cells. The potential of CADPE to the potential of colorectal cancer cells was studied with four inhibitors of MAPK signaling pathway. The effect of CADPE on proliferation inhibition, apoptosis and cell cycle related proteins in human colon cancer was further studied in this experiment. The effect of CADPE on human normal colon cancer tissue cells was evaluated. Experimental results showed that CADPE could significantly inhibit the proliferation and colony formation ability of human colorectal cancer cells and induce the formation of colorectal cancer cells. Cell cycle arrest and apoptosis in G0/G1 phase of colon cancer cells; the inhibition of CADPE on colorectal cancer is related to the activation of p38 MAPK, p53, p21 and p16 signaling pathways, and the inhibition of nuclear factor kappa B and its downstream target gene signal transducer and transcription activator 3 (STAT3) and c-Myc; CADPE can also down regulate survivin survivin and cell weeks The expression of related proteins (such as cyclin D1, CDK4, CDK6, p-Rb and E2F-1); the inhibitors of P38 MAPK can reduce the proliferation inhibition, cell cycle arrest and apoptosis induced by CADPE, but ERK and JNK inhibitors do not have such effects. In the xenograft model of nude mice, the tumor also inhibits tumor growth and induces apoptosis and decreases. The expression of low PCNA, survivin and cell cycle related proteins (cyclin D1, CDK4, CDK6, p-Rb), and CADPE against normal cells in the range of anti-tumor concentration, has high safety in both in vivo and in vitro experiments. Conclusion: experimental results suggest that CADPE plays an important role in the regulation of the proliferation of colorectal cancer cells. It can regulate the proliferation of colon cancer cells through multitarget regulation, induce cell cycle arrest and induce apoptosis of colon cancer cells.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R96
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本文編號(hào)：2062442